GSM7089884: NCI-H1975 + sequins mixA mixed with HCC827 + sequins mixB in ratio 25:75; Homo sapiens; RNA-Seq
The mRNA was extracted using a Qiagen RNA miniprep kit and purified using the NEBNext® Poly(A) mRNA Magnetic Isolation Module (E7490). The NCI-H1975+sequins mixA replicate 1 RNA was mixed with the HCC827+sequins mixB replicate 1 RNA in the 5 following mass ratios: 100:0, 75:25, 50:50, 25:75 and 0:100. 1 ng of each mix was then used for library preparation. The cRNA-PCR Barcoding (SQK-PCS109 with SQK-PBK004) kit was used with the supplied ONT protocol. Amplication for purified mRNA spiked with sequins was done with 16 PCR cycles with a 2 minutes and 30 seconds extension step and a 0.8X final bead clean up. Size selection was performed for the standard ~2kb target. 65 ng of the final library were loaded onto 1 FLO-PRO002 PromethION flow cell (R9.4.1).